Biologische Bundesanstalt für Land- und Forstwirtschaft, D-38104 Braunschweig, Germany, and Center for Microbial Ecology and Department of Microbiology, Michigan State University, East Lansing, Michigan 48824.
Appl Environ Microbiol. 1998 Apr;64(4):1220-5. doi: 10.1128/AEM.64.4.1220-1225.1998.
BIOLOG GN plates are increasingly used to characterize microbial communities by determining the ability of the communities to oxidize various carbon sources. Studies were done to determine whether the BIOLOG GN plate assay accurately reflects the catabolic potential of the inoculum used. To gain insight into which populations of microbial communities contribute to the BIOLOG patterns, denaturing gradient gel electrophoresis and temperature gradient gel electrophoresis (TGGE) were used to assess the diversity of ribotypes in the inocula and individual wells of BIOLOG plates following incubation. These studies were done with microbial communities from the rhizosphere of potatoes and an activated sludge reactor fed with glucose and peptone. TGGE analyses of BIOLOG wells inoculated with cell suspensions from the potato rhizosphere revealed that, compared with the inoculum, there was a decrease in the number of 16S rRNA gene fragments obtained from various wells, as well as a concomitant loss of populations that had been numerically dominant in the inoculum. The dominant fragments in TGGE gels could be assigned to the gamma subclass of the class Proteobacteria, suggesting that fast-growing bacteria adapted to high substrate concentrations were numerically dominant in the wells and may have been primarily responsible for the patterns of substrate use that were observed. Similarly, the community structure changed in wells inoculated with cells from activated sludge; one or more populations were enriched, but all dominant populations of the inoculum could be detected in at least one well. This study showed that carbon source utilization profiles obtained with BIOLOG GN plates do not necessarily reflect the functional potential of the numerically dominant members of the microbial community used as the inoculum.
越来越多的研究采用 BIOLOG GN 板来通过测定微生物群落氧化各种碳源的能力来对其进行特征描述。本研究旨在确定 BIOLOG GN 板测定法是否能准确反映接种物的代谢潜能。为了深入了解哪些微生物群落能够对 BIOLOG 图谱产生影响，本研究采用变性梯度凝胶电泳和温度梯度凝胶电泳（TGGE）来评估接种物和 BIOLOG 板中各个孔的微生物群落的多样性。本研究采用了来自土豆根际的微生物群落和以葡萄糖和蛋白胨为食的活性污泥反应器中的微生物群落进行研究。对土豆根际悬浮液接种的 BIOLOG 孔进行 TGGE 分析表明，与接种物相比，从各个孔中获得的 16S rRNA 基因片段数量减少，同时，在接种物中数量占优势的种群数量减少。在 TGGE 凝胶中发现的优势片段可被分配到 Proteobacteria 类的 gamma 亚类，这表明能够适应高底物浓度的快速生长细菌在数量上占优势，可能是观察到的底物利用模式的主要原因。同样，在接种活性污泥细胞的孔中，群落结构发生了变化；一个或多个种群得到了富集，但接种物中所有的优势种群都可以在至少一个孔中检测到。本研究表明，BIOLOG GN 板获得的碳源利用图谱不一定反映作为接种物的微生物群落中数量占优势的成员的功能潜能。
